Adipose Tissue Engineering: Three Different Approaches to Seed Preadipocytes on a Collagen-Elastin Matrix

Millions of plastic and reconstructive surgical procedures are performed each year to repair soft-tissue defects that result from significant burns, tumor resections, or congenital defects. Tissue-engineering strategies have been investigated to develop methods for generating soft-tissue. Preadipocytes represent a promising autologous cell source for adipose tissue engineering. These immature precursor cells, which are located between the mature adipocytes in the adipose tissue, are much more resistant to mechanical stress and ischemic conditions than mature adipocytes. To use preadipocytes for tissue-engineering purposes, cells were isolated from human adipose tissue and seeded onto scaffolds. Once processed, preadipocytes become subject to the human tissue act and require handling under much tighter regulations. Therefore, we intended to identify any influence caused by processing of preadipocytes prior to seeding on the reconstructed adipose tissue formation. Human preadipocytes were isolated from subcutaneous adipose tissue obtained from discarded tissue during abdominoplasties of healthy men and women. Preadipocytes were divided into 3 groups. Cells of group I were seeded onto the scaffold directly after isolation, cells of group II were proliferated for 4 days before seeding, and cells of group III were proliferated and induced to differentiate before seeded onto the scaffold. A 3-dimensional scaffold (Matriderm, Dr. Otto Suwelack Skin and Health Care GmbH, Billerbeck, Germany) containing bovine collagen and elastin served as a carrier. Fourteen days after isolation, all scaffolds were histologically evaluated, using hematoxylin and eosin, anti-Ki-67 antibody, as well as immunofluorescence labeling with Pref-1 antibody (DLK (C-19), peroxisome proliferator-activated receptor gamma antibody, and DAPI (4',6-diamidino-2-phenylindole). Cells of all groups adhered to the scaffolds on day 21 after isolation. Cells of groups I (freshly isolated preadipocytes) and II (proliferated preadipocytes) adhered well and penetrated into deeper layers of the matrix. In group III (induced preadipocytes), penetration of cells was primarily observed to the surface area of the scaffold. : The collagen-elastin matrix serves as a useful scaffold for adipose tissue engineering. Freshly isolated preadipocytes as well as proliferated preadipocytes showed good penetration into deeper layers of the scaffold, whereas induced preadipocytes attached primarily to the surface of the m